zika virus stocks  (ATCC)

Journal: Viruses

Article Title: An Examination of the Long-Term Neurodevelopmental Impact of Prenatal Zika Virus Infection in a Rat Model Using a High Resolution, Longitudinal MRI Approach

doi: 10.3390/v13061123

Figure Lengend Snippet: Absolute whole-brain volumes in postnatal day 2 neonates across all three groups stratified by sex using both automated and manual methods of segmentation ( n = 8–10 per sex per group). ( A ) Automated segmentation revealed a main effect of inoculation ( # p < 0.05). Specifically, whole-brain volume was significantly smaller in iZIKV pups when compared to both mock control and ZIKV pups (post-hoc, # p < 0.05). Analysis also revealed a main effect of sex (* p < 0.05); P2 females across all groups had significantly smaller whole-brain volumes than their male counterparts. No interaction between inoculation and sex was detected. ( B ) Manual segmentation also revealed a main effect of inoculation ( # p < 0.05). iZIKV pups again displayed significantly reduced whole-brain volumes when compared with all other groups; however, ZIKV pups also displayed significantly reduced whole-brain volumes when compared to mock controls (post-hoc, # p < 0.05). There was no main effect of sex or interaction of inoculation × sex. Bars represent the mean ± SD and data points represent individual subjects. Data were analyzed using a two-way ANOVA with inoculation ( # p < 0.05) and sex (* p < 0.05) as factors, followed by Tukey’s test.

Article Snippet: Zika virus stocks (strain PRVABC059, Puerto Rico, Human, December 2015 from ATCC) were propagated in Vero cells and serially passaged to yield high-titer virus stocks.

Techniques: Control

Journal: Viruses

Article Title: An Examination of the Long-Term Neurodevelopmental Impact of Prenatal Zika Virus Infection in a Rat Model Using a High Resolution, Longitudinal MRI Approach

doi: 10.3390/v13061123

Figure Lengend Snippet: Absolute whole-brain volumes in postnatal day 16 juveniles across all three groups stratified by sex using both automated and manual methods of segmentation ( n = 9–10 per sex per group). ( A ) Automated segmentation revealed a main effect of inoculation ( # p < 0.05). Specifically, whole-brain volume was significantly larger in ZIKV juveniles when compared to the mock control group (post-hoc, # p < 0.05). ( B ) Manual segmentation also revealed a main effect of inoculation ( # p < 0.05). Both iZIKV and ZIKV juveniles displayed significantly reduced whole-brain volumes when compared to mock control juveniles (post-hoc, # p < 0.05 ). Analyses also revealed a main effect of sex for both segmentation methods (* p < 0.05). More specifically, P16 females across all groups had significantly smaller whole-brain volumes than their male counterparts. There was no interaction of inoculation × sex within either method of segmentation. Bars represent the mean ± SD and data points represent individual subjects. Data were analyzed using a two-way ANOVA with inoculation ( # p < 0.05) and sex (* p < 0.05) as factors, followed by Tukey’s test.

Article Snippet: Zika virus stocks (strain PRVABC059, Puerto Rico, Human, December 2015 from ATCC) were propagated in Vero cells and serially passaged to yield high-titer virus stocks.

Techniques: Control

Journal: Viruses

Article Title: An Examination of the Long-Term Neurodevelopmental Impact of Prenatal Zika Virus Infection in a Rat Model Using a High Resolution, Longitudinal MRI Approach

doi: 10.3390/v13061123

Figure Lengend Snippet: Absolute whole-brain volumes in postnatal day 24 juveniles across all three groups stratified by sex using both methods of segmentation ( n = 7–10 per sex per group). ( A ) Automated segmentation revealed no main effect of inoculation. ( B ) Manual segmentation revealed a main effect of inoculation ( # p < 0.05). ZIKV juveniles displayed significantly reduced whole-brain volumes when compared to mock control juveniles (post-hoc, # p < 0.05). Analyses also revealed a main effect of sex for both segmentation methods (* p < 0.05). More specifically, P24 females across all groups had significantly smaller whole-brain volumes than their male counterparts. There was no interaction of inoculation × sex within either method of segmentation. Bars represent the mean ± SD and data points represent individual subjects. Data were analyzed using a two-way ANOVA with inoculation ( # p < 0.05) and sex (* p < 0.05) as factors, followed by Tukey’s test.

Article Snippet: Zika virus stocks (strain PRVABC059, Puerto Rico, Human, December 2015 from ATCC) were propagated in Vero cells and serially passaged to yield high-titer virus stocks.

Techniques: Control

Journal: Viruses

Article Title: An Examination of the Long-Term Neurodevelopmental Impact of Prenatal Zika Virus Infection in a Rat Model Using a High Resolution, Longitudinal MRI Approach

doi: 10.3390/v13061123

Figure Lengend Snippet: ( A ) Automated segmentation of brain regions at postnatal day 2 revealed a significant effect of inoculation. Analysis revealed ZIKV-inoculated neonates had a smaller isocortex when compared to mock-inoculated neonates (post-hoc, # p < 0.05). Furthermore, analysis revealed a significant effect of sex in both the isocortex and cerebellum ( * p < 0.05). Analysis detected no interaction of inoculation × sex. ( B ) Manual segmentation of brain regions at postnatal day 2 revealed a significant effect of inoculation. Within the isocortex, iZIKV-inoculated neonates displayed smaller regional volumes when compared to both mock and ZIKV counterparts (post-hoc, # p < 0.05). Analysis also revealed iZIKV neonates had smaller cerebellums in comparison to the mock controls (post-hoc, # p < 0.05). Analysis detected no significant effect of sex or interaction of inoculation × sex. Bars represent the mean ± SD and data points represent individual subjects ( n = 8–10 per sex per group). Data were analyzed using a two-way ANOVA with inoculation ( # p < 0.05) and sex ( * p < 0.05) as factors, followed by Tukey’s test.

Article Snippet: Zika virus stocks (strain PRVABC059, Puerto Rico, Human, December 2015 from ATCC) were propagated in Vero cells and serially passaged to yield high-titer virus stocks.

Techniques: Comparison

Journal: Viruses

Article Title: An Examination of the Long-Term Neurodevelopmental Impact of Prenatal Zika Virus Infection in a Rat Model Using a High Resolution, Longitudinal MRI Approach

doi: 10.3390/v13061123

Figure Lengend Snippet: ( A ) Automated segmentation of brain regions at postnatal day 16 revealed a significant effect of inoculation. Within the hippocampus region, ZIKV-inoculated juveniles displayed smaller regional volumes when compared to both mock and iZIKV counterparts (post-hoc, # p < 0.05). Within the isocortex and cerebellum, both iZIKV-inoculated and ZIKV-inoculated juveniles displayed smaller regional volumes when compared to mock controls (post-hoc, # p < 0.05). Analysis detected no significant effect of sex or interaction of inoculation × sex. ( B ) Manual segmentation of brain regions at postnatal day 16 revealed a significant effect of inoculation. Within the hippocampus region, ZIKV-inoculated juveniles displayed larger regional volumes when compared to both mock and iZIKV counterparts (post-hoc, # p < 0.05). Within the isocortex, iZIKV-inoculated juveniles displayed smaller regional volumes when compared to both mock and ZIKV counterparts (post-hoc, # p < 0.05). Analysis detected no significant effect of sex or interaction of inoculation × sex. Bars represent the mean ± SD and data points represent individual subjects ( n = 9–10 per sex per group). Data were analyzed using a two-way ANOVA with inoculation ( # p < 0.05) and sex as factors, followed by Tukey’s test.

Article Snippet: Zika virus stocks (strain PRVABC059, Puerto Rico, Human, December 2015 from ATCC) were propagated in Vero cells and serially passaged to yield high-titer virus stocks.

Techniques:

Journal: Viruses

Article Title: An Examination of the Long-Term Neurodevelopmental Impact of Prenatal Zika Virus Infection in a Rat Model Using a High Resolution, Longitudinal MRI Approach

doi: 10.3390/v13061123

Figure Lengend Snippet: ( A ) Automated segmentation of brain regions for postnatal day 24 revealed a main effect of sex in all regions analyzed ( * p < 0.05). Furthermore, analysis identified no significant effect of inoculation or interaction of inoculation × sex. ( B ) Manual segmentation of brain regions at postnatal day 24 revealed a significant effect of inoculation. Within the cerebellum region, ZIKV-inoculated juveniles displayed smaller regional volumes when compared to the iZIKV-inoculated juveniles (post-hoc, # p < 0.05). Analysis detected no significant effect of sex or interaction of inoculation × sex. Bars represent the mean ± SD and data points represent individual subjects ( n = 7–10 per sex per group). Data were analyzed using a two-way ANOVA with inoculation ( # p < 0.05) and sex ( * p < 0.05) as factors, followed by Tukey’s test.

Article Snippet: Zika virus stocks (strain PRVABC059, Puerto Rico, Human, December 2015 from ATCC) were propagated in Vero cells and serially passaged to yield high-titer virus stocks.

Techniques:

Journal: Viruses

Article Title: An Examination of the Long-Term Neurodevelopmental Impact of Prenatal Zika Virus Infection in a Rat Model Using a High Resolution, Longitudinal MRI Approach

doi: 10.3390/v13061123

Figure Lengend Snippet: ( A ) Automated segmentation of brain regions at postnatal day 60 revealed a significant effect of inoculation. Within the hippocampus region, ZIKV-inoculated juveniles displayed smaller regional volumes when compared to the iZIKV-inoculated juveniles (post-hoc, # p < 0.05). Moreover, analysis also revealed a main effect of sex within the hippocampal region ( * p < 0.05). Analysis detected no interaction of inoculation × sex. ( B ) Analysis of manually segmented brain regions for postnatal day 60 revealed no significant effect of inoculation, sex, or interaction of inoculation × sex. Bars represent the mean ± SD and data points represent individual subjects ( n = 7–10 per sex per group). Data were analyzed using a two-way ANOVA with inoculation ( # p < 0.05) and sex ( * p < 0.05) as factors, followed by Tukey’s test.

Article Snippet: Zika virus stocks (strain PRVABC059, Puerto Rico, Human, December 2015 from ATCC) were propagated in Vero cells and serially passaged to yield high-titer virus stocks.

Techniques:

Journal: Viruses

Article Title: An Examination of the Long-Term Neurodevelopmental Impact of Prenatal Zika Virus Infection in a Rat Model Using a High Resolution, Longitudinal MRI Approach

doi: 10.3390/v13061123

Figure Lengend Snippet: Analysis of manually segmented hippocampal subregions for postnatal day 2 ( n = 17–19 per group) ( A ), postnatal day 16 ( n = 18–20 per group) ( B ), postnatal day 24 ( n = 15–20 per group) ( C ), and postnatal day 60 ( n = 16–19 per group) ( D ). At P2, volumetric analysis of hippocampal subregions revealed a significant effect of inoculation within the CA3 of the hippocampus. Specifically, the ZIKV-inoculated pups displayed a significant volume reduction in the CA3 subregion when compared to mock counterparts (post-hoc, * p < 0.05). At P16, analysis revealed a significant effect of inoculation within the CA1 of the hippocampus and dentate gyrus. Specifically, ZIKV-inoculated juveniles displayed significantly larger volumes in both the CA1 subregion and dentate gyrus when compared to mock controls (post-hoc, * p < 0.05 ) . Across all groups, bars represent the mean ± SD in each subregion of the hippocampus while data points represent each individual rat.

Article Snippet: Zika virus stocks (strain PRVABC059, Puerto Rico, Human, December 2015 from ATCC) were propagated in Vero cells and serially passaged to yield high-titer virus stocks.

Techniques:

Journal: Nature biomedical engineering

Article Title: Integrated pipeline for the accelerated discovery of antiviral antibody therapeutics

doi: 10.1038/s41551-020-0594-x

Figure Lengend Snippet: a, Flow cytometric identification of target-specific memory B cells after labeling of magnetically enriched total B cells with ZIKV E antigen. Target-specific B cells were labeled with biotinylated recombinant E protein and detected by fluorochrome-conjugated streptavidin as indicated. Green arrow indicates FACS-sorted cells. Representative data of four experiments is shown. b, Bioinformatics filtering steps to select mAb sequences for synthesis. Timeline to identify sequences of target mAbs and obtain cDNA constructs for recombinant mAb expression is indicated with blue arrow in the timeline chart.

Article Snippet: To identify cells suitable for production of high-titer ZIKV stocks in an antigen-independent manner, we used qRT-PCR to rapidly detect ZIKV viral RNA in a panel of immortalized cell lines including those commonly used for virus propagation Vero-E6 (ATCC), BHK (ATCC), JEG-3 (ATCC), HeLa (ATCC), HEK-293T (ATCC), U20S (ATCC), A549 (ATCC), Huh7, Huh7.5, EA.Hy926 (Horizon), and Hap-1 cell lines.

Techniques: Labeling, Recombinant, Construct, Expressing

Journal: Nature biomedical engineering

Article Title: Integrated pipeline for the accelerated discovery of antiviral antibody therapeutics

doi: 10.1038/s41551-020-0594-x

Figure Lengend Snippet: a, Micro-scale expressed and purified mAbs. Dots indicate average concentration of individual mAbs from assay duplicates, and median mAb yield is shown with horizontal line. LOD – limit of the detection. b, Relationships between binding and neutralizing activities of individual mAbs of the panel shown as a heatmap. Binding to E protein was determined by ELISA and neutralizing activity was measured using RTCA from mAbs purified as in (a) . c, ZIKV E protein binding by potently neutralizing mAbs representing three distinct epitope binding groups. MAb rRSV-90, which is specific to the unrelated respiratory syncytial virus (RSV) fusion protein (F) antigen, served as a control. d, Neutralization of ZIKV Brazil or Dakar strain viruses by representative cross-neutralizing mAbs from (c) , as determined using RTCA. Data shown indicate the mean ± SD of assay triplicates in (c, d) , and represent at least two independent experiments. Timeline to identify mAb candidates for in vivo testing is indicated with a blue arrow in the timeline chart.

Article Snippet: To identify cells suitable for production of high-titer ZIKV stocks in an antigen-independent manner, we used qRT-PCR to rapidly detect ZIKV viral RNA in a panel of immortalized cell lines including those commonly used for virus propagation Vero-E6 (ATCC), BHK (ATCC), JEG-3 (ATCC), HeLa (ATCC), HEK-293T (ATCC), U20S (ATCC), A549 (ATCC), Huh7, Huh7.5, EA.Hy926 (Horizon), and Hap-1 cell lines.

Techniques: Purification, Concentration Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Activity Assay, Protein Binding, Virus, Control, Neutralization, In Vivo

Journal: Nature biomedical engineering

Article Title: Integrated pipeline for the accelerated discovery of antiviral antibody therapeutics

doi: 10.1038/s41551-020-0594-x

Figure Lengend Snippet: a, Serum concentration of human mAbs (2 dpi) that was determined by ELISA from mice that were treated prophylactically (d-1) with indicated mAbs (9 μg/mouse) and challenged with ZIKV (d0). Dots show measurements from individual mice. ZIKV-609, −624, −635, −668, −869, −980, −1006, and FLU5J8 were examined using n= 5 mice per group in one experiment; ZIKV-752 (n=14 mice), −893 (n=11 mice), −940 (n=9 mice), and −117 (n=12 mice) were examined in three independent experiments and for which cumulative data from three experiments are shown. Mean ± SD values are shown. N/A – not available. b, Serum ZIKV titers (2 dpi) from mice treated and challenged as in (a). Dots show measurements from individual mice. n=5 mice per group were examined and data represent one experiment except for ZIKV-752 and −117 treatment groups, where total n=14 mice per group were examined in three independent experiments and for which cumulative data from three experiments are shown. Mean ± SD values are shown. c, Survival of B6 mice that were treated and challenged as in (a) . Overall difference in survival between the groups was estimated using a two-sided log-rank (Mantel cox) test. P values are indicated. d, Low dose (4.5 μg/mouse) prophylaxis of three lead candidate mAbs ZIKV-752, −893, and −940, representing each of three competition-binding groups. n=5 mice per group were examined in one experiment. Mice were treated and challenged as in (a) . Overall difference in survival between the groups was estimated using a two-sided log-rank (Mantel cox) test. P values are indicated. e, Low-dose (9 μg/mouse) post-exposure therapeutic efficacy (1 dpi) of three lead candidate mAbs in mice challenged with ZIKV. n=5 mice per group were examined in one experiment. Overall difference in survival between the groups was estimated using a two-sided log-rank (Mantel cox) test. P values are indicated. The mAbs ZIKV-117 and FLU-5J8 or PBS served as controls. Timeline to scale-up mAb protein production and identify the lead protective mAbs is indicated with blue arrows in the timeline chart.

Article Snippet: To identify cells suitable for production of high-titer ZIKV stocks in an antigen-independent manner, we used qRT-PCR to rapidly detect ZIKV viral RNA in a panel of immortalized cell lines including those commonly used for virus propagation Vero-E6 (ATCC), BHK (ATCC), JEG-3 (ATCC), HeLa (ATCC), HEK-293T (ATCC), U20S (ATCC), A549 (ATCC), Huh7, Huh7.5, EA.Hy926 (Horizon), and Hap-1 cell lines.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Binding Assay, Drug discovery

Journal: Nature biomedical engineering

Article Title: Integrated pipeline for the accelerated discovery of antiviral antibody therapeutics

doi: 10.1038/s41551-020-0594-x

Figure Lengend Snippet: a, Serum concentration of human mAbs (2 dpi) from mice that were treated with the indicated mAb-encoding RNA formulations (40 μg/mouse) on day −1 and challenged with ZIKV on day 0 was determined by ELISA. Dots show measurements from individual mice. n=5 mice per group were examined and data represent one experiment except for ZIKV-117 where n=10 mice per group were examined. Mean ± SD values are shown. N/A – not available. b, Serum ZIKV titers (2 dpi) from mice treated and challenged as in (a) was determined by qRT-PCR. Dots show measurements from individual mice. n=5 mice per group were examined in one experiment except for ZIKV-117 where n=10 mice per group were examined. Mean ± SD values are shown. c, Survival of mice that were treated and challenged as in (a) . n=5 mice per group were examined in one experiment. Overall difference in survival between the groups was estimated using a two-sided log-rank (Mantel cox) test. P values are indicated. The mAbs ZIKV-117 and FLU-5J8 served as controls. Timeline to scale-up mAb-encoding RNA production and identify the lead protective mAb-encoding RNA formulations is indicated with blue arrows in the timeline chart.

Article Snippet: To identify cells suitable for production of high-titer ZIKV stocks in an antigen-independent manner, we used qRT-PCR to rapidly detect ZIKV viral RNA in a panel of immortalized cell lines including those commonly used for virus propagation Vero-E6 (ATCC), BHK (ATCC), JEG-3 (ATCC), HeLa (ATCC), HEK-293T (ATCC), U20S (ATCC), A549 (ATCC), Huh7, Huh7.5, EA.Hy926 (Horizon), and Hap-1 cell lines.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Nature biomedical engineering

Article Title: Integrated pipeline for the accelerated discovery of antiviral antibody therapeutics

doi: 10.1038/s41551-020-0594-x

Figure Lengend Snippet: Summary of binding and functional activities of 20 characterized lead mAbs.

Article Snippet: To identify cells suitable for production of high-titer ZIKV stocks in an antigen-independent manner, we used qRT-PCR to rapidly detect ZIKV viral RNA in a panel of immortalized cell lines including those commonly used for virus propagation Vero-E6 (ATCC), BHK (ATCC), JEG-3 (ATCC), HeLa (ATCC), HEK-293T (ATCC), U20S (ATCC), A549 (ATCC), Huh7, Huh7.5, EA.Hy926 (Horizon), and Hap-1 cell lines.

Techniques: Binding Assay, Functional Assay, Cell Isolation, Enzyme-linked Immunosorbent Assay, Recombinant, Neutralization

Journal: Nature biomedical engineering

Article Title: Integrated pipeline for the accelerated discovery of antiviral antibody therapeutics

doi: 10.1038/s41551-020-0594-x

Figure Lengend Snippet: Animals received one 10 mg/kg dose of mAb ZIKV-117 (n=3 NHPs per group) or mAb PGT121(n=3 NHPs per group) served as a contemporaneous control intravenously on day −1 and then challenged subcutaneously with a target dose of 10 3 PFU of ZIKV strain Brazil the next day. a, qRT-PCR measurement of viremia in plasma and other compartments at indicated time points after virus challenge. b, concentration of ZIKV-117 human mAb that was determined in serum of treated and control NHPs at indicated time points after virus challenge. Data in a,b represent a single experiment.

Article Snippet: To identify cells suitable for production of high-titer ZIKV stocks in an antigen-independent manner, we used qRT-PCR to rapidly detect ZIKV viral RNA in a panel of immortalized cell lines including those commonly used for virus propagation Vero-E6 (ATCC), BHK (ATCC), JEG-3 (ATCC), HeLa (ATCC), HEK-293T (ATCC), U20S (ATCC), A549 (ATCC), Huh7, Huh7.5, EA.Hy926 (Horizon), and Hap-1 cell lines.

Techniques: Control, Quantitative RT-PCR, Clinical Proteomics, Virus, Concentration Assay